LOCUS       sequence-357871-        5694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mouse Nav1.6 knockin with 3xV5 at the C-terminal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5694)
  AUTHORS   Liu H, Wang HG, Pitt GS, Liu ZJ
  TITLE     Direct Observation of Compartment-Specific Localization and Dynamics
            of Voltage-Gated Sodium Channels.
  JOURNAL   J Neurosci. 2022 Jun 6;42(28):5482-98. doi: 
            10.1523/JNEUROSCI.0086-22.2022.
  PUBMED    35672149
REFERENCE   2  (bases 1 to 5694)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1523/JNEUROSCI.0086-22"; journalName: "J Neurosci"; date: 
            "2022-06-6- 6"; volume: "42"; issue: "28"; pages: "5482-98"
COMMENT     Sequence Label: PX552-mScn8a-3xV5-EF1-smFP-flag
FEATURES             Location/Qualifiers
     source          1..5694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        145..393
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     145..165
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     316..335
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        414..489
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             544..585
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             601..642
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             658..699
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     promoter        854..1065
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1078..1346
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      1366..1375
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1375..1398
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1402..1425
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1429..1452
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1981..2004
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2011..2034
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2047..2070
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2077..2100
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2311..2334
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2338..2361
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2365..2388
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2441..2917
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2640..2659)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2957..3097
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2957..3086
                     /label=AAV2 ITR
     rep_origin      3172..3627
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3259..3278)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3469..3490
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3644..3663)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3763..3785
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3823..3841)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3909..4013
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4014..4082,4083..4874)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4232..4251)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5045..5633
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5534..5553
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgagggc ctatttccca tgattccttc atatttgcat
      181 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
      241 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
      301 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
      361 gctttatata tcttgtggaa aggacgaaac accccgacaa ggagaagcag caggttttag
      421 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
      481 agtcggtgct ttttttctag taccggtccg ctgctgcttc tccttgtcgg ttctggatcg
      541 ggtgggaagc ctatacccaa tcctctcctg gggttggaca gcaccggcag tggtagcggt
      601 ggcaagccca tccccaaccc cctgctgggc ctggactcca ccggctccgg cagtggtgga
      661 aaacctattc ccaacccact gctgggactg gattctacag gctcaggtag cggcccgctg
      721 ctgcttctcc ttgtcggtct agaccaagga tctgcgatcg ctccggtgcc cgtcagtggg
      781 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattcgctcc
      841 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
      901 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
      961 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     1021 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg
     1081 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
     1141 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
     1201 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
     1261 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg
     1321 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg gtaccgccac catggactac
     1381 aaggacgacg acgataaagg agattacaaa gatgacgatg ataagggcga ttataaggac
     1441 gacgatgaca agggcggcgt gtcgaaaggc gaggagctgt tcacaggcgt ggtccccatc
     1501 ctggttgagc tggacggtga cgttaacggc cacaaattta gcgtacgtgg tgagggcgag
     1561 ggtgacgcta ccaacggaaa gctgaccctg aaattcatct gcacaactgg caagctgccg
     1621 gtgccctggc ccacgctggt aaccactctg ggcggaggcg ttcaatgttt cagtcgttac
     1681 ccggatcaca tgaagcagca tgacttcttc aagtcggcca tgcccgaagg ctacgtgcag
     1741 gagcgcacca tcagttttaa ggatgacggc acctacaaga ctcgcgccga ggtcaagttc
     1801 gaaggagaca ccctggttaa tcgtatcgag ctcaaaggca tcgatttcaa ggaggacgga
     1861 aatattctgg gccacaagct ggagtacaat tttaacagcc acaatgttta cattactgcc
     1921 gacaagcaga agaacggcat caaggctaat ttcaagattc gtcacaatgt ggagggaggc
     1981 gactataagg atgacgatga caaacaacaa gactataagg acgatgatga caaaggtcag
     2041 caaggcgatt acaaggacga tgacgataag cagcaagatt acaaggatga tgatgataag
     2101 ggaggcgacg gaagcgtgca gctggcagat cattaccaac agaacacgcc cattggtgac
     2161 ggtccagtcc tgctgcctga taaccactac ttgtccaccc agtcggtgct gtcgaaggac
     2221 ccgaacgaga agcgtgacca catggtgctg ctggagttcg tgaccgccgc aggaatcacc
     2281 ctcggcatgg atgagcttta caagggcgga gattataaag atgacgacga caagggcgac
     2341 tacaaagatg atgatgacaa gggtgactac aaagacgacg atgataaata agaattcgat
     2401 atcaagctta tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc
     2461 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
     2521 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
     2581 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     2641 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     2701 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
     2761 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
     2821 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
     2881 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg
     2941 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3001 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3061 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     3121 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     3181 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3241 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3301 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3361 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3421 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3481 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     3541 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3601 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     3661 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3721 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3781 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3841 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3901 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3961 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4021 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4081 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4141 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4201 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4261 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4321 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4381 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4441 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4501 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4561 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4621 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4681 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4741 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4801 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4861 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4921 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4981 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5041 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5101 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5161 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5221 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5281 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5341 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5401 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5461 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5521 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5581 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5641 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
